Use of NK-1 receptor antogonists for treating substance use disorders

ABSTRACT

The present invention provides a method for the treatment or prevention of substance use disorders using an orally active, long acting, CNS-penetrant NK-1 receptor antagonist and pharmaceutical compositions comprising such a NK-1 receptor antagonist.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. §119(a) from GreatBritain Application No. 9625051.9, filed Dec. 2, 1996, Great BritainApplication No. 9701459.1, filed Jan. 24, 1997, Great BritainApplication No. 9713715.2, filed Jun. 27, 1997, and Great BritainApplication No. 9717097.1, filed Aug. 12, 1997

This invention relates to the treatment or prevention of certainsubstance use disorders by the administration of a specific class ofNK-1 receptor antagonists.

A common, though not inevitable, outcome of long-term drug use is theoccurrence of a withdrawal response, which occurs either when the drugis withdrawn or when an antagonist is given and which usually takes theform of some sort of adverse reaction.

A withdrawal syndrome occurs in opiate addicts when the opiate iswithdrawn or when an antagonist, such as naloxone, is given. Thesymptoms consist of yawning, rhinorrhoea, and sweating, followed by theso-called cold turkey, in which there is shivering and goose flesh.Later, nausea, vomiting, diarrhoea, and hypertension may occur. Theacute syndrome subsides within a week, but the-addict may have anxietyand sleep disturbances for several weeks or months after. This syndromecan be avoided by introducing increasing doses of methadone as theopiate is withdrawn, as the later withdrawal of methadone, which has amuch longer duration of action than morphine, may not result in thissyndrome.

Delirium tremens may occur on withdrawal of alcohol from chronicalcoholics. This syndrome consists of disorientation and visualhallucinations.

Withdrawal of benzodiazepines after long-term therapy may result in adisturbance of sleep pattern (rebound insomnia assicated with abnormalsleep patterns), agitation, restlessness, and occasionally epilepticconvulsions.

The pharmacological management of drug withdrawal disorders is notalways effective and is often hampered by the addictive potential ofmany of the drugs which are prescribed to treat withdrawal symptoms.

Neurokinin 1 (NK-1; substance P) receptor antagonists are beingdeveloped for the treatment of a number of physiological disordersassociated with an excess or imbalance of tachykinins, and in particularsubstance P. Examples of conditions in which substance P has beenimplicated include disorders of the central nervous system such asanxiety, depression and psychosis (see, for instance, International(PCT) patent specification Nos. WO 95/16679, WO 95/18124 and WO95/23798).

More recently, International (PCT) patent specification No. WO 96/24353(published Aug. 15, 1996) suggests that a more efficacious and safetreatment of psychiatric disorders would be achieved using a combinationof a tachykinin antagonist and a serotonin agonist or selectiveserotonin reuptake inhibitor (SSRI). However, such a regimen would notbe free of side-effects due to the serotonin agonist or SSRI.

It has been reported that central (i.c.v.) injection of RP67580 couldattenuate naloxone-precipitated withdrawal signs in rats receivingmorphine for five days (Maldonado et al, Neurosci. Letts., 156: 135,1993). This study resembles an earlier report using guinea-pigs showingthat the morphine withdrawal response was inhibited by i.c.v. injectionof peptide substance P antagonists (Johnston & Chahl, Arch. Pharmacol.,343: 283, 1991). These investigators subsequently attempted to block thewithdrawal response in guinea-pigs using systemic administration ofCP-96,345, and found that only high doses (20 mg/kg) were active, andthis could therefore not be attributed with confidence to a specificblockage of NK-1 receptors (Chahl & Johnston, Regul. Pept. Suppl. 1,S43, 1992).

NK-1 receptor antagonists are described in published European PatentSpecification Nos. 0 360 390, 0 394 989, 0 429 366, 0 443 132, 0 482539, 0 512 901, 0 512 902, 0 514 273, 0 514 275, 0 517 589, 0 520 555, 0522 808, 0 528 495, 0 532 456, 0 533 280, 0 536 817, 0 545 478, 0 577394, 0 590 152, 0 599 538, 0 610 793, 0 634 402, 0 686 629, 0 693 489, 0694 535, 0 699 655, 0 699 674, 0 707 006, 0 708 101, 0 714 891, 0 723959, 0 733 632 and 0 776 893; and in International Patent SpecificationNos. 90/05525, 90/05729, 91/09844, 91/18899, 92/01688, 92/06079,92/12151, 92/15585, 92/17449, 92/20661, 92/20676, 92/21677, 93/00330,93/00331, 93/01159, 93/01165, 93/01169, 93/01170, 93/06099, 93/09116,93/10073, 93/14113, 93/18023, 93/19064, 93/21155, 9321181, 93/23380,93/24465, 94/01402, 94/02461, 94/03429, 94/03445, 94/04494, 94/04496,94/05625, 94/07843, 94/10165, 94/10167, 94/10168, 94/10170, 94/11368,94/13639, 94/13663, 94/14767, 94/15903, 94/19320, 94/19323, 94/20500,94/26735, 94/26740, 94/29309, 95/02595, 95/04040, 95/04042, 95/06645,95/07886, 95/07908, 95/08549, 95/11880, 95/14017, 95/15311, 95/16679,95/17382, 95/18124, 95/18129, 95/19344, 95/20575, 95/21819, 96/22525,95/23798, 95/26338, 95/28418, 95/30674, 95/30687, 96/05193, 96/05203,96/06094, 96/07649, 96/10562, 96/16939, 96/18643, 96/20197, 96/21661,96/29304, 96/29317, 96/29326, 96/29328, 96/31214, 96/32385, 96/37489,97/01553, 97/01554, 97/03066, 97/08144, 97/14671, 97/17362, 97/18206,97/19084, 97/19942 and 97/21702; and in British-Patent SpecificationNos. 2 266 529, 2 268 931, 2 269 170, 2 269 590, 2 271 774, 2 292 144, 2293 168, 2 293 169, and 2 302 689.

In view of the short-comings of existing therapy for substance usedisorders, there is a need for new, safe and effective treatment forsuch disorders.

The present invention provides the use of a CNS penetrant NK-1 receptorantagonist in an oral, once-a-day medicament for the treatment ofsubstance use disorders. The compounds of this class advantageouslyexhibit a rapid onset of action and a reduced side-effect profile whencompared against conventional therapy.

In particular, the present invention provides a means for theidentification of NK-1 receptor antagonists which would be effective inan oral once-a-day medicament for the treatment of substance usedisorders. The aforementioned patent specifications which describe NK-1receptor antagonists provide no reliable method for the identificationof such compounds.

The exceptional pharmacology of the class of NK-1 receptor antagonistsof use in the present invention enables the treatment of substance usedisorders, without the need for concomitant therapy.

Furthermore, the exceptional pharmacology of the class of NK-1 receptorantagonists of use in the present invention results in a rapid onset ofaction.

The present invention accordingly provides the use of an orally active,long acting, CNS-penetrant NK-1 receptor antagonist (as hereinafterdefined) for the manufacture of a medicament adapted for oraladministration for the treatment or prevention of substance usedisorders.

The present invention also provides a method for the treatment orprevention of substance use disorders, which method comprises the oraladministration to a patient in need of such treatment of an effectiveamount of an orally active, long acting, CNS-penetrant NK-1 receptorantagonist (as hereinafter defined).

In a further aspect of the present invention, there is provided an oralpharmaceutical composition for the treatment of substance use disorderswhich comprises an orally active, long acting, CNS-penetrant NK-1receptor antagonist (as hereinafter defined), together with apharmaceutically acceptable carrier or excipient.

As used herein, the term "substance use disorders" includes substancedependence or abuse with or without physiological dependence. Thesubstances associated with these disorders are: alcohol, amphetamines(or amphetamine-like substances), caffeine, cannabis, cocaine,hallucinogens, inhalants, nicotine, opioids, phencyclidine (orphencyclidine-like compounds), sedative-hypnotics or benzodiazepines,and other (or unknown) substances and combinations of all of the above.

In particular, the term "substance use disorders" includes drugwithdrawal disorders such as alcohol withdrawal with or withoutperceptual disturbances; alcohol withdrawal delirium; amphetaminewithdrawal; cocaine withdrawal; nicotine withdrawal; opioid withdrawal;sedative, hypnotic or anxiolytic withdrawal with or without perceptualdisturbances; sedative, hypnotic or anxiolytic withdrawal delirium; andwithdrawal symptoms due to other substances. It will be appreciated thatreference to treatment of nicotine withdrawal includes the treatment ofsymptoms associated with smoking cessation.

Other "substance use disorders" include substance-induced anxietydisorder with onset during withdrawal; substance-induced mood disorderwith onset during withdrawal; and substance-induced sleep disorder withonset during withdrawal.

As used herein, the term "treatment" refers both to the preventionand/or acute and maintenance treatment of the aforementioned conditions.

Preferred NK-1 receptor antagonists for use in the present invention areselected from the classes of compounds described in European PatentSpecification No. 0 577 394, and International Patent Specification Nos.95/08549, 95/18124, 95/23798 and 96/05181, and International PatentApplication No. PCT/GB97/01630. The preparation of such compounds isfully described in the aforementioned publications.

Particularly preferred NK-1 receptor antagonists of use in the presentinvention include:

2-(S)-(3,5-bis(trifluoromethyl)benzyloxy)-3(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)-3-(S)-phenyl-morpholine;

2-(S)-(3,5-bis(trifluoromethyl)benzyloxy)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)-3-(S)-phenyl-morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(5-(N,N-dimethylamino)methyl-1,2,3-triazol-4-yl)methyl-3-(S)-phenylmorpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(5-(N,N-dimethylamino)methyl-1,2,3-triazol-4-yl)methyl-3-(S)-(4-fluorophenyl)morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(4-monophosphoryl-5-oxo-1H-1,2,4-triazolo)methyl)morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(1-monophosphoryl-5-oxo-1H-1,2,4-triazolo)methyl)morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(2-monophosphoryl-5-oxo-1H-1,2,4-triazolo)methyl)morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxyphosphoryl-1H-1,2,4-triazolo)methyl)morpholine;

2-(S)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(1-monophosphoryl-5-oxo-4H-1,2,4-triazolo)methyl)morpholine;

2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(4-N,N-dimethylaminobut-2-yn-yl)-3-(S)-(4-fluorophenyl)morpholine;

(3S,5R,6S)-3-2-cyclopropoxy-5-(trifluoromethoxy)phenyl-1-oxa-7-aza-spiro 4.5!decane;

(3R,5R,6S)-3-2-cyclopropoxy-5-(trifluoromethoxy)phenyl-1-oxa-7-aza-spiro 4.5!decane;

or a pharmaceutically acceptable salt thereof.

Full descriptions of the preparation of the NK-1 receptor antagonistswhich may be employed in the present invention may be found in thereferences cited herein.

Suitable pharmaceutically acceptable salts of the NK-1 receptorantagonists of use in the present invention include acid addition saltswhich may, for example, be formed by mixing a solution of the compoundwith a solution of a pharmaceutically acceptable non-toxic acid such ashydrochloric acid, fumaric acid, maleic acid, succinic acid, aceticacid, citric acid, tartaric acid, carbonic acid, phosphoric acid orsulphuric acid. Salts of amine groups may also comprise the quaternaryammonium salts in which the amino nitrogen atom carries an alkyl,alkenyl, alkynyl or aralkyl group. Where the compound carries an acidicgroup, for example a carboxylic acid group, the present invention alsocontemplates salts thereof, preferably non-toxic pharmaceuticallyacceptable salts thereof, such as the sodium, potassium and calciumsalts thereof.

Preferably the compositions containing an NK-1 receptor antagonist ofuse according to the present invention are in unit dosage forms such astablets, pills, capsules, wafers and the like. Additionally, the NK-1receptor antagonists of use according to the present invention may bepresented as granules or powders for extemporaneous formulation asvolume defined solutions or suspensions. Alternatively, the NK-1receptor antagonists of use according to the present invention may bepresented in ready-prepared volume defined solutions or suspensions.Preferred forms are tablets and capsules.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical carrier, e.g. conventionaltableting ingredients such as corn starch, lactose sucrose, sorbitol,talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, andother pharmaceutical diluents, e.g. water, to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention, or a non-toxic pharmaceuticallyacceptable salt thereof. When referring to these preformulationcompositions as homogeneous, it is meant that the active ingredient isdispersed evenly throughout the composition so that the composition maybe readily subdivided into equally effective unit dosage forms such astablets, pills and capsules. This solid preformulation composition isthen subdivided into unit dosage forms of the type described abovecontaining from 0.1 to about 500 mg of the active ingredient of thepresent invention. The tablets or pills of the novel composition can becoated or otherwise compounded to provide a dosage form affording theadvantage of prolonged action. For example, the tablet or pill cancomprise an inner dosage and an outer dosage component, the latter beingin the form of an envelope over the former. The two components can beseparated by an enteric layer which serves to resist disintegration inthe stomach and permits the inner component to pass intact into theduodenum or to be delayed in release. A variety of materials can be usedfor such enteric layers or coatings, such materials including a numberof polymeric acids and mixtures of polymeric acids with such materialsas shellac, cetyl alcohol and cellulose acetate.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally include aqueoussolutions, suitably flavoured syrups, aqueous or oil suspensions, andflavoured emulsions with edible oils such as cottonseed oil, sesame oil,coconut oil, peanut oil or soybean oil, as well as elixirs and similarpharmaceutical vehicles. Suitable dispersing or suspending agents foraqueous suspensions include synthetic and natural gums such astragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose,methylcellulose, polyvinyl-pyrrolidone or gelatin.

Compositions of the present invention may also be administered via thebuccal cavity using conventional technology, for example, absorptionwafers.

Compositions in the form of tablets, pills, capsules or wafers for oraladministration are particularly preferred.

A minimum dosage level for the NK-1 receptor antagonist is about 5 mgper day, preferably about 10 mg per day and especially about 20 mg perday. A maximum dosage level for the NK-1 receptor antagonist is about1500 mg per day, preferably about 1000 mg per day and especially about500 mg per day. The compounds are administered once a day.

It will be appreciated that the amount of the NK-1 receptor antagonistrequired for use in the treatment or prevention of substance usedisorders will vary not only with the particular compounds orcompositions selected but also with the route of administration, thenature of the condition being treated, and the age and condition of thepatient, and will ultimately be at the discretion of the patient'sphysician or pharmacist.

Two compounds of use in the present invention which are described inInternational Patent Application No. PCT/GB97/01630 may be preparedaccording to the following methods:

PREPARATION 1

(2S)-1-tert-Butoxycarbonyl-2-phenylpiperidin-3-one

Dimethyl sulfoxide (20.80 ml, 22.90 g, 29.3 mmol) in dichloromethane (75ml) was added dropwise to a cooled (-70° C.) solution of oxalyl chloride(13.95 ml, 20.30 g, 160 mmol) in dichloromethane (350 ml). The mixturewas stirred at -70° C. for 15 minutes, then(2S,3S)-1-tert-butoxycarbonyl-3-hydroxy-2-phenylpiperidine (prepared bythe method described in European Patent Specification number 0 528495-A; 36.91 g, 133 mmol) in dichloromethane (150 ml) was addeddropwise. The mixture was stirred at -70° C. for 20 minutes, thenallowed to warm to -30° C. The mixture was cooled to -50° C. andtriethylamine (55.95 ml, 40.45 g, 400 mmol) was added slowly. Themixture was allowed to warm to 0° C. and diluted with ice-cooleddichloromethane (250 ml). The mixture was washed with ice cold aqueouscitric acid solution (5%, 2×300 ml) and water (300 ml), dried (MgSO₄),and the solvent was evaporated under reduced pressure to give the titlecompound as a yellow oil (42.3 g), which was used immediately withoutfurther purification. ¹ H NMR (250 MHz, CDCl₃) δ 7.5-7.3 (5H, m), 5.8(1H, br s), 4.2 (1H, br s), 3.4 (1H, m), 2.6 (2H, m), 2.0 (2H, m), and1.54 (9H, s).

PREPARATION 2

(2S,3R)-1-tert-Butoxycarbonyl-3-hydroxy-3-(2-methylene-3-phenoxypropyl)-2-phenylpiperidine

A solution of 3-(chloromagnesio)-2-(phenoxymethyl)-1-propene in THF(0.91M, 3 ml) (Louw et. al., Tetrahedron, 48, 6087-6104, 1992, preparedfrom 2.74 mmol of 3-chloro-2-(phenoxymethyl)-1-propene) was slowly addedto a solution of (2S)-1-tert-butoxycarbonyl-2-phenylpiperidin-3-one(Preparation 1) in THF (3 ml). The mixture was stirred at roomtemperature for 1 hours, then saturated aqueous ammonium chloride (20ml) was added and the mixture was extracted with ethyl acetate (20 ml).The organic phase was washed with brine, dried (MgSO₄) and the solventwas evaporated under reduced pressure. The residue was purified bycolumn chromatography on silica gel, eluting with hexane/ethyl acetate(100:0 increasing to 80:20) to give the title compound. ¹ H NMR (360MHz, CDCl₃) δ 7.48 (2H, d, J=6.9 Hz), 7.35-7.2 (6H, m), 6.9-6.88 (3H,m), 5.4 (1H, s), 5.15 (2H, d, J=13.7 Hz), 4.61 (2H, s), 4.11 (2H, m),3.17 (1H, m), 2.66 and 2.59 (2H, AB d, J=14.0 Hz), 1.95 (2H, m), 1.79(2H, m), and 1.36 (9H, s). m/z (ES⁺) 424 (M+1).

PREPARATION 3

(5R,6S)-3-Methylene-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

To a cooled (-80° C.) solution of(2S,3R)-1-tert-butoxycarbonyl-3-hydroxy-3-(2-methylene-3-phenoxypropyl)-2-phenylpiperidine(Preparation 2, 1.53 g, 3.62 mmol) in THF (20 ml) was added n-butyllithium (2.5M in hexanes, 1.45 ml, 3.62 mmol) followed by a solution ofzinc chloride (0.5M in THF, 7.24 ml, 3.62 mmol). The solution wasallowed to warm to room temperature andtetrakis(triphenylphosphine)palladium (0) (0.23 g, 0.2 mmol) was added.The mixture was degassed with bubbling nitrogen and heated under refluxfor 16 hours. The mixture was cooled and the solvent was evaporatedunder reduced pressure. The residue was partitioned between ethylacetate and 2M sodium hydroxide. The organic phase was washed withsaturated brine, dried (MgSO₄) and purified by chromatography on acolumn containing silica gel (eluting with hexane containing increasingproportions of ethyl acetate between 0% to 5%). Evaporation of thefractions gave(6S,5R)-3-methylene-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane. ¹ H NMR (360 MHz, CDCl₃) δ 7.58 (2H, d, J=8.4 Hz), 7.32-7.21(3H, m), 5.23 (1H, s), 5.06 (1H, m), 4.97 (1H, m), 4.39 (2H, AB d,J=13.3 Hz), 3.99 (1H, dd, J=13.3, 4.48 Hz), 2. 83 (1H, ABd J=15.5 Hz),2.7 (1H,td J=12.5, 3.93 Hz), 2.5 (1H, ABd, J=15.4 Hz), 2.15 (2H, td,J=12., 0.4 Hz), 1.69 (2H, m), and 1.46 (9H,s). m/z (ES⁺) 329 (M+2H-^(t)BuOCO).

PREPARATION 4

(5R,6S)-3-Keto-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

Through a cooled (-80° C.) solution of(5R,6S)-3-methylene-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 3; 0.665 g) in dichloromethane (5 ml) andmethanol (5 ml) was bubbled a mixture of ozone and oxygen for 45minutes. After the solution had been purged with nitrogen, dimethylsulphide (0.5 ml) was added and then stirred under nitrogen at roomtemperature for 16 hours. The solvent was removed in vacuo and theresidue partitioned between ethyl acetate and water. The organic phasewas dried (MgSO₄), evaporated and the residue purified by chromatographyon a column containing silica gel (eluting with hexane containingincreasing proportions of ethyl acetate between 0% to 10%). Evaporationof the fractions gave the title compound. ¹ H NMR (250 MHz, CDCl₃) δ7.58 (2H, d, J=6.2 Hz), 7.37-7.26 (3H, m), 5.3 (1H, s), 4.15 and 4.09(2H, AB d, J=17.4 Hz), 3.97 (1H, m), 2.80 (1H, td, J=12.9, 4.0 Hz), 2.74and 2.48 (2H, ABd, J=18.1 Hz), 2.29 (2H, m), 1.88-1.63 (2H, m), and 1.44(9H, s). m/z (ES⁺) 332 (M+1).

PREPARATION 5(5R,6S)-3-Trifluoromethylsulfonyloxy-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!dec-3-ene

To a cooled (-80° C.) solution of 1M sodium hexamethyldisilazide (0.38ml, 0.38 mmol) in THF was added a solution of(5R,6S)-3-keto-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 4; 0.105 mg, 0.319 mmol) in THF (3 ml). Thesolution was stirred for 1 hours at -80° C. then a solution of 2-N,N-bis(trifluoromethylsulfonyl)amino!-5-chloropyridine (0.163 g, 0.415mmol) in THF (3 ml) was added. The solution was stirred at -80° C. for30 minutes then at room temperature for 30 minutes before being quenchedby addition of saturated ammonium chloride solution and ethyl acetate.The dried (MgSO₄) organic phase was purified by chromatography on acolumn containing silica gel (eluting with hexane containing increasingproportions of ethyl acetate between 0% to 5%). Evaporation of thefractions gave the title compound. ¹ H NMR (360 MHz, CDCl₃) δ 7.4 (2H,d, J=7.3 Hz), 7.3-7.22 (3H, m), 6.01 (1H, t, J=2.13 Hz), 5.13 (1H, s),4.56 and 4.26 (2H, ABdd, J=12.4, 1.97 Hz), 4.10 (1H, dt, J=12.6, 4.22Hz), 3.00 (1H, m), 2.28-2.04 (2H, m), 1.88-1.76 (2H, m), and 1.37 (9H,s). m/z (ES⁺) 464 (M+1).

PREPARATION 6

(5R,6S)-3-Trimethylstannyl-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!dec-3-ene

To a degassed solution of(5R,6S)-3-trifluoromethylsulfonyloxy-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!dec-3-ene (Preparation 5; 0.482 g, 1.04 mmol), lithium chloride(0.264 g, 6.25 mmol), lithium carbonate (0.076 g) and hexamethyldistannane (0.96 g, 2.9 mmol) in THF (10 ml) was addedtriphenylphosphine palladium (0) (0.06 g). The solution was degassed andthen heated at 60° C. for 5 hours under nitrogen. Water (20 ml) andethyl acetate (20 ml) were added and the dried organic phase waspurified by chromatography on a column containing silica gel (elutingwith hexane containing increasing proportions of ethyl acetate between0% to 5%). Evaporation of the fractions gave the title compound as acrystalline solid. ¹ H NMR (360 MHz, CDCl₃) δ 7.25 (2H, d, J=7.3 Hz),7.1-7.0 (3H, m), 5.83 (1H, t, J=2.5 Hz), 4.78 (1H, s), 4.48 and 4.02(2H, dd, J=12.9, 2.3 Hz), 3.96 (1H, dd, J=6.16, 13.4 Hz), 2.95 (1H, td,J=13.3, 4.5 Hz), 1.84 (1H, m), 1.68 (1H, m), 1.60 (2H, m), 1.19 (9H, s),and 0.0 (6H, s).

PREPARATION 7

(2S,3R)-1-tert-Butoxycarbonyl-3-(3-hydroxypropyn-1-yl)-2-phenylpiperidin-3-ol

O-Trimethylsilylpropargyl alcohol (24.51 ml, 20.47 g, 160 ml) was addedslowly to a cooled (-10° C.) solution of ethylmagnesium bromide (1M intetrahydrofuran, 160 ml, 160 mmol). The mixture was stirred at 0° C. for20 minutes, then at room temperature for 2 hours. The mixture was cooledto -10° C. and a solution of(2S)-1-tert-butoxycarbonyl-2-phenylpiperidin-3-one (Preparation 1; 42.3g) in tetrahydrofuran (200 ml) was added dropwise over 30 minutes.(Internal temperature below -5° C.). The mixture was stirred at roomtemperature for 14 hours, poured into water (300 ml) and saturatedaqueous ammonium chloride (300 ml) and extracted with ethyl acetate(2×300 ml). The combined organic fractions were washed with brine (300ml), dried (MgSO₄) and the solvent was evaporated under reducedpressure. The residue was dissolved in ethyl acetate (500 ml) and asolution of tetrabutylammonium fluoride (1M in THF, 160 ml, 160 mmol)was added dropwise. The mixture was stirred at room temperature for 30minutes, water (300 ml) was added, and the layers were separated. Theaqueous layer was extracted with ethyl acetate (2×300 ml) and thecombined organic fractions were washed with water (300 ml) and brine(300 ml), dried (MgSO₄) and the solvent was evaporated under reducedpressure to give the crude title compound as an orange oil (45 g). Thecrude material was purified by flash column chromatography on silicagel, eluting with hexane/ethyl acetate (90:10 increasing to 25:75) togive the title compound as an amber oil (32.2 g). ¹ H NMR (CDCl₃) δ7.53-7.55 (2H, m), 7.19-7.35 (3H, m), 5.56 (1H, s), 4.27 (2H, s),3.99-4.03 (1H, m), 3.25 (1H, br s), 2.77-2.81 (1H, m), 2.77 (1H, br s),2.12-2.20 (1H, m), 1.91-1.99 (2H, m), 1.77-1.83 (1H, m), and 1.39 (9H,s).

PREPARATION 8

2-Bromo-4-(trifluoromethoxy)phenol

To a cooled (0° C.) solution of 4-trifluoromethoxyphenol (35.6 g, 0.2mol) in chloroform (280 ml) was added dropwise a solution of bromine (32g, 0.2 mol) in chloroform (50 ml). The solution was stirred at 0° C. for1 hour and at room temperature for 2 hours. Dichloromethane (200 ml) andwater (400 ml) ware added and the organic phase was washed further withwater (400 ml), brine (200 ml) and dried (MgSO₄). The solvent wasremoved and the residue was purified by distillation at reduced pressureto give the title compound. ¹ H NMR (250 MHz, CDCl₃) δ 7.38 (1H, d,J=2.1 Hz), 7.13 (1H, dd, J=9.1, 2.1 Hz), 7.03 (1H, d, J=9.1 Hz), and5.53 (1H, s).

PREPARATION 9

2-Benzyloxy-5-(trifluoromethoxy)bromobenzene

2-Bromo-4-(trifluoromethoxy)phenol (Preparation 8; 5 g, 20 mmol) wasdissolved in N,N-dimethylformamide (60 ml), and potassium carbonate (5.4g, 40 mmol) was added, followed by benzyl bromide (3.5 ml, 30 mmol), andthe reaction was stirred at ambient temperature for 15 hours. Thereaction was diluted with water (150 ml) and extracted into ethylacetate (3×60 ml). The combined organic fractions were washed with water(100 ml), brine (100 ml), dried (MgSO₄) and evaporated in vacuo.Purification on silica, eluting with 2% and 5% ethyl acetate in hexanegave the title compound as a clear oil (6.7 g, 96%). ¹ H NMR (250 MHz,CDCl₃) δ 5.47 (2H, s), 7.23 (1H, d, J=9 Hz), 7.43 (1H, dd J=8.2, 2.9Hz), and 7.75 (6H, m).

PREPARATION 10

Z-(2S,3R)-1-tert-Butoxycarbonyl-3-(3-hydroxyprop-1-en-1-yl)-2-phenylpiperidin-3-ol

Palladium on calcium carbonate, poisoned with lead (Lindlar catalyst, 2g) was added to a solution of(2S,3R)-1-tert-butoxycarbonyl-3-(3-hydroxypropyn-1yl)-2-phenylpiperidin-3-ol(Preparation 7; 32 g, 96.6 mmol) in ethyl acetate (300 ml) and themixture was stirred under hydrogen (1 atmosphere) for 4 hours. Themixture was filtered and the solvent was evaporated under reducedpressure to give the title compound as an oil (32 g, 100%). ¹ H NMR (360MHz, CDCl₃) δ 7.42 (2H, d, J=7.6 Hz), 7.35-7.25 (3H, m), 5.83 (1H, d,J12.3 Hz), 5.68 (1H, dt, J=12.3, 6.0 Hz), 5.06 (1H, s), 4.27 (1H, m),4.12 (2H, m), 3.32 (1H, m), 3.13 (1H, s), 2.28 (1H, t, J=5.9 Hz), 2.02(1H, m), 1.92-1.78 (3H, m), and 1.32 (9H, s). m/z (ES⁺) 334 (M+1).

PREPARATION 11

(5R,6S)-6-Phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro 4.5!dec-3-ene

Diethylazodicarboxylate (18.2 ml, 115 mmol) in THF (100 ml) was addeddropwise to a solution ofZ-(2S,3R)-1-tert-butoxycarbonyl-3-(3-hydroxyprop-1-en-1-yl)-2-phenylpiperidin-3-ol(Preparation 10; 32 g, 96 mmol) and triphenylphosphine (30.2 g, 115mmol) in THF (700 ml). The mixture was stirred at 0° C. for 30 minutesthen at room temperature for 1.5 hours. The solvent was evaporated underreduced pressure and the residue was purified by flash columnchromatography on silica gel, eluting with hexane/ethyl acetate (95:5increasing to 80:20) to give the title compound as a colorless solid(23.4 g, 77%). ¹ H NMR (CDCl₃) δ 7.45 (2H, d, J=7.4 Hz), 7.27 (2H, t,J=7.4 Hz), 7.20 (1H, t, J=7.4 Hz), 6.03 (1H, dt, J=6.1, 2.0 Hz), 5.68(1H, dt, J=6.1, 2.0 Hz), 5.06 (1H, s), 4.61 (1H, dt, J=13.1, 2.0 Hz),4.32 (1H, dt, J=13.1, 2.0 Hz), 4.08 (1H, m), 3.05 (1H, m), 2.05 (1H, m),1.75 (3H, m), and 1.37 (9H, s). m/z (ES⁺) 316 (M+1).

PREPARATION 12

2-Benzyloxy-5-(trifluoromethoxy)benzene

Benzyl bromide (66.17 ml, 95.35 g, 0.56 mol) was added to a mixture of4-(trifluoromethoxy)phenol (90.26 g, 0.51 mol) and potassium carbonate(140.97 g, 1.2 mol) in dimethylformamide (160 ml) and the mixture wasstirred at room temperature for 72 hours. The mixture was poured intowater (1.5 l) and extracted with ethyl acetate (3×50 ml). The combinedorganic fractions were washed with aqueous sodium carbonate (saturated,500 ml), dried (MgSO₄) and the solvent was evaporated under reducedpressure to give the title compound as a colorless solid (133.5 g, 99%).¹ H NMR (360 MHz, CDCl₃) δ 7.39 (5H, m), 7.14 (2H, d, J=9.0 Hz), 6.95(2H, d, J=9.0 Hz), and 5.05 (2H, s).

PREPARATION 13

2-Benzyloxy-5-(trifluoromethoxy)iodobenzene

Iodine (71.96 g, 0.28 mol) in chloroform was added dropwise to a mixtureof 2-benzyloxy-5-(trifluoromethoxy)benzene (Preparation 12, 73.06 g,0.27 mol) and silver trifluoroacetate (71.57 g, 0.32 mol) indichloromethane and the mixture was stirred at room temperature for 18hours. The mixture was filtered through celite, washed with aqueoussodium thiosulfate (5%, 2×2 l), dried (MgSO₄) and the solvent wasevaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with hexane/ethyl acetate,to give the title compound as a colorless oil (108.03 g), containing 11%unreacted 2-benzyloxy-5-(trifluoromethoxy)iodobenzene. ¹ H NMR (360 MHz,CDCl₃) δ 7.67 (1H, d, J=2.8 Hz), 7.40 (5H, m), 7.16 (1H, dd, J=8.9, 2.8Hz), 6.82 (1H, d, J=8.9 Hz), and 5.14 (2H, s).

PREPARATION 14

(5R,6S)-3-(2-Benzyloxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!dec-3-ene

(5R,6S)-3-Trimethylstannyl-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!dec-3-ene(Preparation 6; 6.43 mmol), lithium chloride (0.163 g),benzyloxy-5-(trifluoromethoxy)phenol (Preparation 9; 7.7 mmol) intoluene (25 ml) was degassed before addition of triphenyiphosphinepalladium (0) (0.37 g). The solution was degassed thoroughly beforeheating to 110° C. for 14 hours. The solution was partitioned betweenwater and ethyl acetate and the dried organic phase was purified bychromatography on a column containing silica gel (eluting with hexanecontaining increasing proportions of ethyl acetate between 0% to 4%) togive the title compound. ¹ H NMR (360 MHz, CDCl₃) δ 1.33 (9H, s), 1.65(1H, m), 1.76 (2H, m), 2.08 (1H, m), 3.11 (1H, m), 4.08 (1H, m), 4.60(1H, dd, J=12.2 Hz, J=2 Hz), 4.92 (1H, dd, J=12.1 Hz, J=1.8 Hz), 5.08(1H, s), 5.1 (2H, q, J=11.5 Hz), 6.65 (1H, s), 6.94 (2H, d, J=8.9 Hz),7.08 (1H, d, J=9 Hz), 7.18 (2H, t, J=8.1 Hz), 7.25 (3H, m), 7.38 (5H,m).

PREPARATION 15

(3S,5R,6S)-3-(2-Hydroxy-5-trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

(5R,6S)-3-(2-Benzyloxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!dec-3-ene (Preparation 14) (3.88 g) was dissolved in ethyl acetate(15 ml) and methanol (15 ml). Palladium hydroxide on carbon (1.00 g) wasadded and the suspension was shaken under a hydrogen atmosphere (50 psi)for 72 hours. The mixture was filtered and the solvent was evaporatedunder reduced pressure. The residue was purified by medium pressurechromatography on silica gel, eluting with hexane/ethyl acetate (75:25)to give(3R,5R,6S)-3-(2-hydroxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (191 mg), ¹ H NMR (250 MHz, CDCl₃) δ 7.70 (2H, d, J=7.3 Hz),7.33 (2H, t, J=7.3 Hz), 7.26 (1H, d, J=7.3 Hz), 7.05 (1H, br s), 6.96(2H, m), 6.82 (1H, d, J=9.4 Hz), 5.43 (1H, s), 4.27 (1H, m), 4.01 (1H,m), 3.95 (1H, m), 3.73 (1H, m), 2.73 (2H, m), 2.33 (1H, m), 1.87-1.58(4H, m); and 1.50 (9H, s).and(3S,5R,6S)-3-(2-hydroxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (2.3 g), ¹ H NMR (360 MHz, CDCl₃) δ 1.38 (9H, s), 1.73 (2H,m), 1.81 (1H, m), 2.18 (2H, m), 2.50 (1H, m), 2.81 (1H, m), 3.62 (1H, t,J=7.2 Hz), 3.92 (1H, m), 3.98 (1H, d, J=13.2 Hz), 4.23 (1H, m), 5.33(1H, s), 6.75 (1H, d, J=8.5 Hz), 6.94 (2H, m), 7.25 (1H, m), 7.31 (2H,m), and 7.55 (2H, d, J=7.8 Hz).

PREPARATION 16

(3R,5R,6S)-3-(2-Benzyloxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

A mixture of 2-benzyloxy-5-(trifluoromethoxy)iodobenzene (Preparation13, 21.8 g, 55.2 mmol),(5R,6S)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro 4.5!dec-3-ene(Preparation 11, 7.0 g, 22.1 mmol), tetra-n-butylammonium chloride (6.18g, 22.2 mmol), lithium chloride (9.35 g, 0.22 mol) and potassium formate(5.64 g, 67.0 mmol) in dimethylformamide (100 ml) was degassed with afirestone valve (5 x). Palladium acetate (491 mg, 2.2 mmol) was addedand the mixture was degassed with a firestone valve (5 x). The mixturewas stirred at 60° C. for 15 hours, then further2-benzyloxy-5-(trifluoromethoxy)iodobenzene (Preparation 13, 4.32 g,11.0 mmol), potassium formate (2.78 g, 33.5 mmol) and palladium acetate(260 mg, 1.1 mmol) were added. The mixture was stirred at 60° C. for 22hours, cooled and filtered. The solvent was evaporated under reducedpressure, water (600 ml) was added and the mixture was extracted withethyl acetate (2×300 ml). The combined organic fractions were washedwith brine (300 ml), dried (MgSO₄) and the solvent was evaporated underreduced pressure. The residue was purified by flash columnchromatography on silica gel, eluting with hexane/dichloromethane (75:25increasing to 0:100) then dichloromethane/ethyl acetate (95:5), to givethe title compound (9.42 g, 73%). ¹ H NMR (360 MHz, CDCl₃) δ 7.56 (2H,d, J=7.7 Hz), 7.40-7.20 (8H, m), 7.14 (1H, d, J=2.0 Hz), 7.00 (1H, dd,J=8.9, 2.0 Hz), 6.88 (1H, d, J=8.9 Hz), 5.30 (1H, s), 5.08 (2H, s), 4.27(1H, m), 3.97 (1H, m), 3.87 (2H, m), 2.78 (1H, m), 2.56 (1H, m), 2.15(1H, m), 1.96 (1H, m), 1.67 (3H, m), and 1.42 (9H, s).

PREPARATION 17

(3R,5R,6S)-3-(2-Hydroxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

Palladium on carbon (10%, 0.59 g) was added to a solution of(3R,5R,6S)-3-(2-benzyloxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 16, 6.10 g, 10.5 mmol) in methanol-water (99:1,200 ml) and the mixture was stirred under hydrogen (50 psi.) for 72hours. The mixture was filtered, washing with ethanol, and the solventwas evaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with dichloromethane/ethylacetate (99:1 increasing to 90:10) to give the title compound. ¹ H NMR(360 MHz, CDCl₃) 6 7.70 (2H, d, J=7.3 Hz), 7.33 (2H, t, J=7.3 Hz), 7.26(1H, d, J=7.3 Hz), 7.05 (1H, br s), 6.96 (2H, m), 6.82 (1H, d, J=9.4Hz), 5.43 (1H, s), 4.27 (1H, m), 4.01 (1H, m), 3.95 (1H, m), 3.73 (1H,m), 2.73 (2H, m), 2.33 (1H, m), 1.87-1.58 (4H, m), and 1.50 (9H, s).

PREPARATION 18

(3S,5R,6S)-3-2-(1-Phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

(3S,5R,6S)-3-(2-Hydroxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 15) (290 mg, 0.59 mmol) was dissolved in toluene(5 ml) and silver carbonate (179 mg, 0.65 mmol) was added in oneportion. (1-Iodocycloprop-1-yl)phenylsulfide (Cohen T. and Matz J. R.,J. Am. Chem. Soc. 1980, 102, 6902) (180 mg, 0.65 mmol) was then addedover one minute at room temperature. The mixture was stirred at 55° C.for 4 hours, then further portions of silver carbonate (179 mg, 0.65mmol) and (1-iodocycloprop-1-yl)phenylsulfide (180 mg, 0.65 mmol) wereadded. The mixture was stirred at 55° C. for a further 3 hours, cooled,filtered and the solvent was evaporated under reduced pressure. Theresidue was purified by column chromatography on silica gel, elutingwith hexane/ethyl acetate (90:10 increasing to 80:20) to give the titlecompound as a colourless oil (120 mg, 32%). ¹ H NMR (250 MHz, CDCl₃) δ7.55-7.44 (4H, m), 7.36-7.23 (7H, m), 7.13-7.02 (2H, m), 5.16 (1H, brs), 4.09 (1H, t, J=6 Hz), 4.03-3.92 (1H, m), 3.67-3.49 (2H, m),2.94-2.79 (1H, m), 2.26 (1H, dd, J=7.9, 12.9 Hz), 2.15-2.01 (2H, m),1.76-1.59 (3H, m), 1.53-1.45 (4H, m), and 1.36 (9H, s). m/z (ES⁺) 642(M+1).

PREPARATION 19

(3R,5R,6S)-3-2-(1-Phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

Prepared from(3R,5R,6S)-3-(2-hydroxy-5-(trifluoromethoxy)phenyl)-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 17) according to the method of Preparation 18. ¹H NMR (360 MHz, CDCl₃) δ 7.57 (2H, app. d, J=7.6 Hz), 7.45 (2H, app. d,J=7.7 Hz), 7.36-7.19 (7H, m), 7.16-7.06 (2H, m), 5.28 (1H, br s), 4.13(1H, app. t, J=7.8 Hz), 3.96 (1H, br. d, J=13 Hz), 3.80-3.60 (2H, m),2.79 (1H, br. t, J=13 Hz), 2.50 (1H, dd, J=13, 7.9 Hz), 2.17 (1H, dt,J=13, 4.6 Hz), 1.80 (1H, dd, J=12, 9.8 Hz), 1.75-1.38 (7H, m), and 1.44(9H, s). m/z (ES⁺) 642 (M+1).

PREPARATION 20

(3S,5R,6S)-3-2-Cyclopropoxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

Naphthalene (120 mg, 0.936 mmol) was dissolved in THF (1.5 ml) undernitrogen and freshly cut lithium metal (7.0 mg, 0.94 mmol) was added.The mixture was then sonicated at room temperature for 20 minutes toproduce a dark green solution of lithium naphthalenide. This solutionwas cooled to -78° C., then (3S,5R,6S)-3-2-(1-phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 18) (120 mg, 0.187 mmol) in THF (0.5 ml) wasadded over 1 minute. The reaction mixture was stirred for 30 minutes,then water (5 ml) and ether (10 ml) were added. The layers wereseparated and the aqueous layer was extracted with ether (10 ml). Thecombined organic fractions were dried (MgSO₄) and the solvent wasevaporated under reduced pressure. The residue was purified by columnchromatography on silica gel, eluting with hexane/ethyl acetate (90:10increasing to 80:20) to give the title compound as a colourless oil(58.6 mg, 59%). ¹ H NMR (250 MHz, CDCl₃) δ 7.58-7.52 (2H, m), 7.36-7.17(4H, m), 7.10-7.01 (2H, m), 5.18 (1H, br s), 4.20 (1H, t, J=6.7 Hz),4.05-3.95 (1H, m), 3.76-3.55 (3H, m), 2.92-2.79 (1H, m), 2.37 (1H, dd,J=12.9, 7.8 Hz), 2.18-2.06 (2H, m), 1.80-1.67 (3H, m), 1.38 (9H, s), and0.86-0.73 (4H, m). m/z (ES⁺) 534 (M+1).

PREPARATION 21

(3R,5R,6S)-3-2-Cyclopropoxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane

Naphthalene (120 mg, 0.936 mmol) was dissolved in THF (1.5 ml) undernitrogen and freshly cut lithium metal (7.0 mg, 0.94 mmol) was added.The mixture was then sonicated at room temperature for 20 minutes toproduce a dark green solution of lithium naphthalenide. A solution of(3R,5R,6S)-3-2-(1-phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 19, 135 mg, 0.21 mmol) in THF (2 ml) undernitrogen was cooled to -78° C. and the solution of lithium naphthalenidein THF was added dropwise until the intense green colour persisted. Thereaction was then stirred for one minute, water (5 ml) was added and themixture was warmed to room temperature. Ether (10 ml) was added and thelayers were separated. The aqueous phase was extracted with a furtherportion of ether (10 ml) and the combined organic phases were dried(MgSO₄) and the solvent was evaporated under reduced pressure. Theresidue was purified by column chromatography on silica gel, elutingwith hexane/ethyl acetate (50:50) to give the title compound as acolourless oil (87 mg, 78%). ¹ H NMR (360 MHz, CDCl₃) δ 7.59 (2H, app.d, J=7.6 Hz), 7.32 (2H, app. t, J=7.6 Hz), 7.27-7.18 (2H, m), 7.11-7.03(2H, m), 5.32 (1H, br s), 4.29-4.21 (1H, m), 3.97 (1H, br. d, J=13 Hz),3.83-3.68 (3H, m), 2.76 (1H, dt, J=13, 4.1 Hz), 2.55 (1H, dd, J=13, 7.2Hz), 2.22 (1H, dt, J=12, 5.2 Hz), 1.85 (1H, dd, J=13, 9.9 Hz), 1.80-1.63(3H, m), 1.46 (9H, s), and 0.82-0.76 (4H, m). m/z (ES⁺) 534 (M+1).

COMPOUND A

(3S,5R,6S)-3-2-Cyclopropoxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-aza-spiro4.5!decane Hydrochloride

Trifluoroacetic acid (2.5 ml) was added dropwise to a stirred, cooled 0°C.) solution of (3S,5R,6S)-3-2-cyclopropoxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-(tert-butoxycarbonyl)aza-spiro4.5!decane (Preparation 20; 492 mg, 0.92 mmol) in dichloromethane (25ml) and the mixture was stirred at room temperature for 3 hours. Themixture was poured into water (50 ml), the pH was adjusted to 10.0 withaqueous sodium hydroxide (4M) and the mixture was extracted withdichloromethane (3×50 ml). The combined organic fractions were dried(MgSO₄) and the solvent was evaporated under reduced pressure. Theresidue was purified by flash column chromatography on silica gel,eluting with dichloromethane/methanol/ammonia (aq.) (96:4:0.4 increasingto 94:6:0.6). The residue was dissolved in ethanol (20 ml), cooled inice and ethereal hydrogen chloride (1M, 1.8 ml, 1.8 mmol) was addeddropwise. The mixture was stirred at 0° C. for 5 minutes, then thesolvent was evaporated under reduced pressure. The residue wascrystallized from ether (20 ml)/ethanol (0.5 ml) and the solid wascollected and dried in vacuo to give the title compound as a colorlesssolid (354 mg, 89%). m.p. 214-216° C., ¹ H NMR (500 MHz, CD₃ OD) δ 7.59(2H, m), 7.52 (3H, m), 7.26 (1H, d, J=8.9 Hz), 7.03 (1H, dd, J=8.9, 2.2Hz), 6.20 (1H, d, J=2.2 Hz), 4.85 (2H, br s), 4.43 (1H, s), 4.19 (1H, t,J=8.0 Hz), 3.87 (1H, quin, J=8.0 Hz), 3.76 (1H, m), 3.44 (1H, m), 3.25(2H, m) 2.29-1.78 (6H, m), 0.80 (2H, m), and 0.66 (2H, m). m/z (ES⁺) 434(M+1). Found: C, 61.41; H, 5.51; N, 3.08. C₂₄ H₂₆ F₃ NO₃.HCl requires:C, 61.34; H, 5.79; N, 2.98%.

COMPOUND B

(3R,5R,6S)-3-2-Cyclopropoxy-5-(trifluoromethoxy)phenyl!-6-phenyl-1-oxa-7-aza-spiro4.5!decane

Prepared from the compound of Preparation 21 according to the methodused for Compound A. ¹ H NMR (360 MHz, CDCl₃) δ 7.50-7.42 (2H, m),7.36-7.26 (3H, m), 7.03 (1H, d, J=8.9 Hz), 6.95 (1H, br. d, J=8.9 Hz),6.81 (1H, br s), 3.92 (1H, t, J=7.4 Hz), 3.62-3.53 (2H, m), 3.50 (1H,s), 3.20 (1H, dd, J=12, 4.2 Hz), 2.77 (1H, dt, J=12, 2.8 Hz), 2.30-1.93(4H, m), 1.87 (1H, br s), 1.71-1.49 (3H, m), 0.76-0.65 (2H, m), and0.65-0.54 (2H, m). m/z (ES⁺) 434 (M+1).

A further compound and diastereomers thereof of use in the presentinvention may be prepared according to the following methods:

DESCRIPTION 1

2-(1-Phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)benzaldehyde

Silver carbonate (1.2 g, 4.34 mmol) was added to a solution of2-hydroxy-5-(trifluoromethoxy)benzaldehyde (0.5 g, 2.43 mmol) and(1-iodocycloprop-1-yl)phenylsulfide (Cohen T. and Matz J. R., J. Am.Chem. Soc. 1980, 102, 6902) (1.2 g, 4.34 mmol) in toluene (30 mL) andthe mixture was stirred at 40° C. overnight. The mixture was cooled,diluted with ethyl acetate and filtered, washing well with ethylacetate. The mixture was washed with aqueous sodium hydroxide, dried(MgSO₄) and the solvent was evaporated under reduced pressure. Theresidue was purified by flash column chromatography on silica gel,eluting with hexane/Et₂ O (95:5), to give the title compound as a yellowoil (191 mg, 27%). ¹ H NMR (360 MHz, CDCl₃) δ 1.51-1.56 (2H, m),1.44-1.48 (2H, m), 7.25-7.35 (7H, m), 7.69 (1H, d, J 2.0 Hz), and 10.26(1H, s).

DESCRIPTION 2

2-Cyclopropoxy-5-(trifluoromethoxy)benzaldehyde

Freshly cut lithium metal (97 mg, 13.9 mmol) was added to a solution ofnaphthalene (1.77 g, 13.9 mmol) in THF (20 mL) and the mixture wassonicated at room temperature for 30 min. to produce a dark greensolution of lithium naphthalenide. A solution of2-(1-phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)benzaldehyde(Description 1, 96 mg, 0.27 mmol) in THF (2 mL) was cooled to -78° C.and the solution of lithium naphthalenide in THF (2 mL) was addeddropwise until the intense green colour persisted. The reaction was thenstirred for 5 min., water (6 mL) was added and the mixture was warmed toroom temperature. The mixture was extracted with ethyl acetate, thecombined organic fractions were dried (MgSO₄) and the solvent wasevaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with hexane/Et₂ O (80:20),to give to give the title compound as a colourless oil (4 mg, 6%). ¹ HNMR (360 MHz, CDCl₃) δ 0.86 (4H, m), 3.82-3.9 (1H, m), 7.42 (2H, m),7.62 (1H, d, J 2.5 Hz), and 10.36 (1H, s).

DESCRIPTION 3

2-Nitro-4-(trifluoromethoxy)phenol

Iron(111)nitrate nonahydrate (1.97 g, 4.87 mmol) was added to a solutionof 4-(trifluoromethoxy)phenol (2 g, 11.24 mmol) in ethanol (20 mL) andthe mixture was heated under reflux overnight. The mixture was allowedto cool to room temperature, acidified to pH 1 with aqueous hydrochloricacid (1M) and extracted with ethyl acetate. The combined organicfractions were dried (MgSO₄), and the solvent was evaporated underreduced pressure. The residue was purified by short columnchromatography on silica gel, eluting with hexane/EtOAc (70:30), to givethe title compound as a yellow oil (2.25 g, 89%). ¹ H NMR (360 MHz,CDCl₃) δ 10.53 (1H, s), 8.01 (1H, d, J 3.0 Hz), 7.49 (1H, dd, J 9.1, 3.0Hz), and 7.23 (1H, d, J 9.1 Hz).

DESCRIPTION 4

2-(1-Phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)nitrobenzene

Prepared from the compound of Description 3 according to the method ofDescription 1. ¹ H NMR (360 MHz, CDCl₃) δ 7.73 (1H, d, J 2.7 Hz), 7.58(1H, d, J 9.2 Hz), 7.50-7.24 (6H, m), 1.57-1.53 (2H, m), and 1.44-1.40(2H, m).

DESCRIPTION 5

2-Cyclopropoxy-5-(trifluoromethoxy)benzeneamine

Prepared from the compound of Description 4 according to the method ofDescription 2. ¹ H NMR (360 MHz, CDCl₃) δ 7.06 (1H, dd, J 2.8, 6.7 Hz),6.56 (2H, m), 3.83 (2H, br s), 3.74 (1H, m), and 0.79 (4H, m). m/z (ES⁺)234 (M+1).

DESCRIPTION 6

2-(1-Phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)benzeneamine

Iron powder (13.5 g, 241 mmol) was added to a suspension of2-(1-phenylthiocycloprop-1-yl)oxy-5-(trifluoromethoxy)nitrobenzene(Description 4, 11.27 g, 30.1 mmol) in water (300 mL) and acetic acid(75 mL) and the mixture was stirred at 80° C. overnight. The mixture wascooled and filtered through celite, washing with ether. The filtrate wasextracted with ether, the combined organic fractions were washed withaqueous sodium hydroxide (1M), dried (MgSO₄), and the solvent wasevaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with hexane/Et₂ O (90:10increasing to 80:20), to give the title compound as a yellow solid (8 g,78%). ¹ H NMR (360 MHz, CDCl₃) δ 7.48 (2H, m), 7.34-7.23 (3H, m), 7.15(1H, d, J 8.74 Hz), 6.60-6.56 (2H, m), 3.78 (2H, br s), 1.49-1.46 (2H,m), and 1.39-1.35 (2H, m).

DESCRIPTION 7

2-Cyclopropoxy-5-(trifluoromethoxy)benzeneamine

Prepared from the compound of Description 6 according to the method ofDescription 2. ¹ H NMR (360 MHz, CDCl₃) δ 7.06 (1H, dd, J 2.8, 6.7 Hz),6.56 (2H, m), 3.83 (2H, br s), 3.74 (1H, m), and 0.79 (4H, m). m/z (ES⁺)234 (M+1).

DESCRIPTION 8

2-Cyclopropoxy-5-(trifluoromethoxy)iodobenzene

An ice-cooled solution of sodium nitrite (3.55 g, 51 mmol) in water (10mL) was added dropwise to a stirred, cooled (0° C.) solution of2-cyclopropoxy-5-(trifluoromethoxy)benzeneamine (Description 7, 4.8 g,20.6 mmol) in aqueous hydrochloric acid (5M, 300 mL), maintaining theinternal temperature at 0° C. The mixture was stirred at 0° C. for 30min., then potassium iodide (8.55 g, 51.5 mmol) in water (10 mL) wasadded dropwise, maintaining the internal temperature at 0° C. Themixture was stirred at 0° C. for 30 min., then allowed to warm up toroom temperature and stirred until nitrogen evolution ceased. Themixture was extracted with ether, the organic fraction was washed withaqueous sodium thiosulfate (10%), dried (MgSO₄), and the solvent wasevaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with hexane/Et₂ O (98:2increasing to 95:5), to give the title compound as a colourless oil(6.23 g, 88%). ¹ H NMR (360 MHz, CDCl₃) δ 7.62 (1H, d, J 2.4 Hz), 7.20(1H, dd, J 9.1, 2.4 Hz), 7.15 (1H, d, J 9.1 Hz), 3.80 (1H, m), and 0.83(4H, m).

DESCRIPTION 9

2-Cyclopropoxy-5-(trifluoromethoxy)benzaldehyde

A solution of 2-cyclopropoxy-5-(trifluoromethoxy)iodobenzene(Description 8, 0.344 g, 1 mmol) in toluene (2.5 mL) was degassed withbubbling nitrogen for 10 min. Tetrakis(triphenylphosphine)palladium (0)(15 mg) was added, the mixture was degassed with bubbling nitrogen for afurther 5 min., then carbon monoxide was bubbled through the mixture for10 min. The mixture was warmed to 50° C. and a solution of tributyl tinhydride (0.3 mL, 1.1 mmol) in toluene (5 mL) was added at a rate of 2mL/h. via a syringe pump, maintaining carbon monoxide bubblingthroughout. The mixture was cooled, diluted with ether (20 mL) andaqueous potassium fluoride solution (50%) was added. The mixture wasstirred at room temperature overnight, filtered and the layers wereseparated. The organic layer was dried (MgSO₄), and the solvent wasevaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with hexane/Et₂ O (80:20),to give the title compound as a colourless oil. 1H NMR (360 MHz, CDCl₃)δ 0.86 (4H, m), 3.82-3.9 (1H, m), 7.42 (2H, m), 7.62 (1H, d, J 2.5 Hz),and 10.36 (1H, s).

DESCRIPTION 10

(±)-(2RS)-1-tert-Butoxycarbonyl-2-phenylpiperidin-3-one

Dimethyl sulfoxide (32.0 mL, 35.3 g, 0.45 mol) in dichloromethane (100mL) was added dropwise to a cooled (-70° C.) solution of oxalyl chloride(18.7 mL, 27.5 g, 0.22 mol) in dichloromethane (1000 mL). The mixturewas stirred at -70° C. for 15 min., then(2S,3S)-1-tert-butoxycarbonyl-3-hydroxy-2-phenylpiperidine (prepared bythe method described in European Patent Specification number 0 528495-A; 50 g, 0.18 mol) in dichloromethane (150 mL) was added dropwise.The mixture was stirred at -70° C. for 1 h., then triethylamine (125.8mL, 91.3 g, 0.9 mol) was added slowly. The mixture was stirred at roomtemperature for 1 h., water (250 mL) and aqueous sodium hydrogencarbonate (saturated, 250 mL) were added and the mixture was stirred atroom temperature overnight. The layers were separated and the aqueouslayer was extracted with dichloromethane (2×300 mL). The combinedorganic fractions were washed with brine, dried (MgSO₄) and the solventwas evaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with hexane/EtOAc (90:10),to give the title compound as a yellow oil (45.0 g, 91%). ¹ H NMR (250MHz, CDCl₃) δ 7.5-7.3 (5H, m), 5.8 (1H, br s), 4.2 (1H, br s), 3.4 (1H,m), 2.6 (2H, m), 2.0 (2H, m), and 1.54 (9H, s).

DESCRIPTION 11

(±)-(2R3R,2S3S)-1-(tert-Butoxycarbonyl)-2-phenylpiperidin-3-amine

A solution of hydroxylamine hydrochloride (17 g, 0.24 mol) and sodiumacetate (55.67 g, 0.41 mol) in water (150 mL) was added to a solution of(±)-(2RS)-1-tert-butoxycarbonyl-2-phenylpiperidin-3-one (Description 10,45 g, 0.16 mol) in ethanol (300 mL) and the mixture was stirred at roomtemperature for 1 h. The solvent was evaporated under reduced pressure,water was added and the mixture was extracted with ethyl acetate. Theorganic fraction was washed with brine, dried (MgSO₄) and the solventwas evaporated under reduced pressure. The residue was dissolved inethanol (400 mL) and Raney nickel (50 g) was added. The mixture wasshaken under hydrogen (40 psi) overnight, filtered and the solvent wasevaporated under reduced pressure. The residue was purified by flashcolumn chromatography on silica gel, eluting with CH₂ Cl₂ /MeOH (100:0increasing to 85:15), to give the title compound as a colorless oil(10.9 g, 24%). ¹ H NMR (360 MHz, CDCl₃) δ 7.43 (2H, d, J 7.0 Hz), 7.30(3H, m), 5.19 (1H, d, J 6.2 Hz), 4.00 (1H, m), 3.17 (2H, m), 1.90-1.64(4H, m), 1.36 (9H, s), and 1.26 (2H, br s).

COMPOUND C (±)-(2R3R,2S3S)-N-{2-Cyclopropoxy-5-(trifluoromethoxy)phenyl!methyl}-2-phenylpiperidin-3-amineDihydrochloride

2-Cyclopropoxy-5-(trifluoromethoxy)benzaldehyde (Description 9, 55 mg,0.21 mmol) was added to(±)-(2R3R,2S3S)-1-(tert-butoxycarbonyl)-2-phenylpiperidin-3-amine(Description 11, 58 mg, 0.21 mmol), citric acid (89 mg, 0.42 mmol) and 3Å molecular sieves in dry methanol (5 mL) and the mixture was stirred atroom temperature for 1.5 h. Sodium borohydride (30 mg) was added and themixture was stirred at room temperature for 2 h. Ethyl acetate was addedand the mixture was washed with aqueous hydrochloric acid (0.1M, 2×25mL) and brine (25 mL), dried (MgSO₄) and the solvent was evaporatedunder reduced pressure. The residue was dissolved in dichloromethane (3mL), cooled to 0° C. and trifluoroacetic acid (2 mL) was added slowly.The mixture was stirred at room temperature for 1 h., the solvent wasevaporated under reduced pressure and ethyl acetate was added. Themixture was washed with aqueous sodium hydrogen carbonate (saturated,2×25 mL) and brine (25 mL), dried (MgSO₄) and the solvent was evaporatedunder reduced pressure. The residue was purified by flash columnchromatography on silica gel, eluting with CH₂ Cl₂ /MeOH/NH₃ (Aq.)(96:4:0.4). The residue was dissolved in ethanol (2 mL), cooled in iceand ethereal hydrogen chloride (1M, 0.24 mL, 0.24 mmol) was added. Thesolvent was evaporated under reduced pressure and the residue wasrecrystallised from ethanol to give the title compound as a colorlesssolid (20 mg, 20%). m.p. 169-171° C. ¹ H NMR (400 MHz, CD₃ OD) δ 0.64(1H, m), 0.80 (3H, m), 1.99 (1H, m), 2.24 (1H, m), 2.46 (2H, m), 3.30(1H, m), 3.64 (1H, m), 3.75 (2H, m), 3.96 (1H, br s), 4.08 (1H, m), 4.95(1H, s), 7.23 (1H, s), 7.31 (1H, d, J 9.0 Hz), 7.37 (1H, d, J 9.0 Hz),7.54 (3H, m), and 7.67 (2H, m). m/z (ES⁺) 407 (M+1).

Particularly preferred NK-1 receptor antagonists of use in the presentinvention are compounds which are potent NK-1 receptor antagonists, i.e.compounds with an NK-1 receptor affinity (IC₅₀) of less than 10 nM,favourably less than 2 nM and preferably less than 1 nM.

The class of orally active, long acting, CNS-penetrant NK-1 receptorantagonists of use in the present invention is identified using acombination of the following assays:

ASSAY 1: NK-1 Receptor binding

NK-1 receptor binding assays are performed in intact Chinese hamsterovary (CHO) cells expressing the human NK-1 receptor using amodification of the assay conditions described by Cascieri et al, J.Pharmacol. Exp. Ther., 1992, 42, 458. The receptor is expressed at alevel of 3×10⁵ receptors per cell. Cells are grown in monolayer culture,detached from the plate with enzyme-free dissociation solution(Speciality Media Inc.), and washed prior to use in the assay. ¹²⁵I-Tyr⁸ -substance P (0.1 nM, 2000 Ci/mmol; New England Nuclear) isincubated in the presence or absence of test compounds (dissolved in 5μl dimethylsulphoxide, DMSO) with 5×10⁴ CHO cells. Ligand binding isperformed in 0.25 ml of 50 mM Tris-HCl, pH7.5, containing 5 mM MnCl₂,150 mM NaCl, 0.02% bovine serum albumin (Sigma), 50 μg/ml chymostatin(Peninsula), 0.1 nM phenylmethylsulphonyl fluoride, 2 μg/ml pepstatin, 2μg/ml leupeptin and 2.8 μg/ml furoyl saccharine. The incubation proceedsat room temperature until equilibrium is achieved (>40 minutes) and thereceptor-ligand complex is harvested by filtration over GF/C filterspre-soaked in 0.1% polyethylenimine using a Tomtek 96-well harvester.Non-specific binding is determined using excess substance P (1 μM) andrepresents <10% of total binding.

ASSAY 2: Gerbil Foot-Tapping

CNS-penetrant NK-1 receptor antagonists for use in the present inventioncan be identified by their ability to inhibit foot tapping in gerbilsinduced by anxiogenic agents (such as pentagastrin) or central infusionof NK-1 receptor agonists such as GR73632, or caused by aversivestimulation such as foot shock or single housing, based on the method ofRupniak & Williams, Eur. J. Pharmacol., 1994, 265, 179.

Briefly, male or female Mongolian gerbils (35-70 g) are anaesthetised byinhalation of an isoflurane/oxygen mixture to permit exposure of thejugular vein in order to permit administration of test compounds orvehicle in an injection volume of 5 ml/kg i.v. Alternatively, testcompounds may be administered orally or by subcutaneous orintraperitoneal routes. A skin incision is then made in the midline ofthe scalp to expose the skull. An anxiogenic agent (e.g. pentagastrin)or a selective NK-1 receptor agonist (e.g. GR73632 (d AlaL-Pro⁹,Me-Leu¹⁰ !-substance P-(7-11)) is infused directly into thecerebral ventricles (e.g. 3 pmol in 5 μl i.c.v., depending on testsubstance) by vertical insertion of a cuffed 27 gauge needle to a depthof 4.5 mm below bregma. The scalp incision is closed and the animalallowed to recover from anaesthesia in a clear perspex observation box(25 cm×20 cm×20 cm). The duration of hind foot tapping is then recordedcontinuously for approximately 5 minutes. Alternatively, the ability oftest compounds to inhibit foot tapping evoked by aversive stimulation,such as foot shock or single housing, may be studied using a similarmethod of quantification.

ASSAY 3: Ferret Emesis

Individually housed male ferrets (1.0-2.5 kg) are dosed orally by gavagewith test compound. Ten minutes later they are fed with approximately100 g of tinned cat food. At 60 minutes following oral dosing, cisplatin(10 mg/kg) is given i.v. via a jugular vein catheter inserted under abrief period of halothane anaesthesia. The catheter is then removed, thejugular vein ligated and the skin incision closed. The ferrets recoverrapidly from the anaesthetic and are mobile within 10-20 minutes. Theanimals are observed continuously during recovery from the anaestheticand for 4 hours following the cisplatin injection. The numbers ofretches and vomits occurring during the 4 hours after cisplatinadministration are recorded by trained observers.

ASSAY 4: Separation-Induced Vocalisation

Male and female guinea-pigs pups are housed in family groups with theirmothers and littermates throughout the study. Experiments are commencedafter weaning when the pups are 2 weeks old. Before entering anexperiment, the pups are screened to ensure that a vigorous vocalisationresponse is reproducibly elicited following maternal separation. Thepups are placed individually in an observation cage (55 cm×39 cm×19 cm)in a room physically isolated from the home cage for 15 minutes and theduration of vocalisation during this baseline period is recorded. Onlyanimals which vocahse for longer than 5 minutes are employed for drugchallenge studies (approximately 50% of available pups may fail to reachthis criterion). On test days each pup receives an oral dose or an s.c.or i.p. injection of test compound or vehicle and is then immediatelyreturned to the home cage with its mother and siblings for 30 to 60minutes before social isolation for 15 minutes as described above. Theduration of vocalisation on drug treatment days is expressed as apercentage of the pre-treatment baseline value for each animal. The samesubjects are retested once weekly for up to 6 weeks. Between 6 and 8animals receive each test compound.

As used herein, the term "CNS-penetrant" refers to NK-1 receptorantagonists which are able to inhibit NK-1 receptor antagonist-inducedfoot-tapping in the gerbil as hereinafter defined.

Essentially, hind foot-tapping in the gerbil induced by infusion of theNK-1 receptor agonist, GR73632 (d Ala L-Pro⁹,Me-Leu¹⁰ !-substanceP-(7-11)), under anaesthesia, directly into the central ventricles isinhibited when a CNS-penetrant NK-1 receptor antagonist is administeredintravenously immediately prior to GR73632 challenge, wherein hindfoot-tapping over a period of five minutes following recovery from theanaesthesia is inhibited with an ID₅₀ ≦3 mg/kg, and preferably with anID₅₀ ≦1 mg/kg.

In an alternative method, the NK-1 receptor antagonist is administeredorally, 1 hour prior to GR73632 challenge, wherein the foot-tapping overa period of five minutes following recovery from anaesthesia isinhibited with an ID₅₀ ≦30 mg/kg, and preferably with an ID₅₀ ≦10 mg/kg.

CNS-penetrant NK-1 receptor antagonists of use in the present inventionare also effective in the attenuation of separation-inducedvocalisations by guinea-pig pups as hereinafter defined.

Essentially, a vocalisation response in guinea-pig pups is induced byisolation from their mothers and littermates, which response isattenuated when a CNS-penetrant NK-1 receptor antagonist is administeredsubcutaneously 30 minutes prior to isolation, wherein vocalisationsduring the first 15 minutes of isolation are attenuated with an ID₅₀ ≦20mg/kg, preferably with an ID₅₀ ≦10 mg/kg, and especially with an ID₅₀ ≦5mg/kg.

In an alternative method, the NK-1 receptor antagonist is administeredorally, 4 hours prior to isolation, wherein vocalisations during thefirst 15 minutes of isolation are attenuated with an ID₅₀ ≦20 mg/kg,preferably with an ID₅₀ ≦10 mg/kg, and especially with an ID₅₀ ≦5 mg/kg.

A suitable selection cascade for NK₁ antagonists of use according to thepresent invention is as follows:

(i) Determine affinity for human NK₁ receptor in radioligand bindingstudies (Assay 1); select compounds with IC₅₀ ≦10 nM, preferably IC₅₀ ≦2nM, especially IC₅₀ ≦1 nM.

(ii) Determine ability of compounds to penetrate CNS by their ability toinhibit foot tapping in gerbils induced by central injection of an NK₁agonist (Assay 2); select compounds that inhibit foot tapping with ID₅₀≦3 mg/kg i.v., and preferably ID₅₀ ≦1 mg/kg i.v. when administeredimmediately prior to central NK₁ agonist challenge, or ID₅₀ ≦30 mg/kgp.o., and preferably ID₅₀ ≦10 mg/kg p.o. 1 hour prior to challenge.

(iii) Determine central duration of action of compounds in gerbil foottapping assay following intravenous administration 24 hours prior tocentral NK₁ agonist challenge; select compounds showing ≦25-fold loss ofpotency compared with ID₅₀ determined in step (ii) above with theproviso that ID₅₀ ≦10 mg/kg i.v., and preferably ≦5 mg/kg i.v. after 24hour pre-treatment.

(iv) Determine oral bioavailability of compounds by pharmacokineticanalysis, activity in gerbil foot tapping assay following oraladministration and/or by ability to inhibit cisplatin-induced emesis inferrets (Assay 3); select compounds with ID₉₀ ≦3 mg/kg p.o., andpreferably ID₉₀ ≦1 mg/kg p.o.

Particularly preferred compounds of use in the present invention areidentified using steps (i) to (iv) followed by step (v):

(v) Determine activity of compounds in assays sensitive to conventionalanxiolytic drugs (inhibition of pharmacologically evoked foot tapping ingerbils and/or inhibition of distress vocalisations in guinea-pig pups(Assay 4)). Select compounds with ID₅₀ ≦20 mg/kg, and preferably ID₅₀≦10 mg/kg.

Yet further preferred compounds of use in the present invention may beselected from those compounds which satisfy the NK-1 receptor bindingcriteria of step (i) which, in addition, have ≦5-fold shift in affinitywhen incubated in the presence of human serum albumin (HSA) to shownon-specific protein binding.

One example of a NK-1 receptor antagonist of use in the presentinvention is the compound2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine,the preparation of which is described in International PatentSpecification No. WO 95/16679. In the aforementioned assays, thiscompound has the following activity:

human NK-1 receptor binding: IC₅₀ =0.1 nM

gerbil foot-tapping (5 mins.): ID₅₀ =0.36 mg/kg i.v.

gerbil foot-tapping (24 hrs.): ID₅₀ =0.33 mg/kg i.v.

ferret emesis: ID₉₀ <3 mg/kg p.o.

The following example illustrates pharmaceutical compositions accordingto the invention.

EXAMPLE 1 Tablets containing 50-300 mg of NK-1 Antagonist

    ______________________________________                  Amount mg    ______________________________________    NK-1 antagonist 50.0       100.0  300.0    Microcrystalline cellulose                    80.0       80.0   80.0    Modified food corn starch                    80.0       80.0   80.0    Lactose         189.5      139.5  139.5    Magnesium Stearate                    0.5        0.5    0.5    ______________________________________

The active ingredient, cellulose, lactose and a portion of the cornstarch are mixed and granulated with 10% corn starch paste. Theresulting granulation is sieved, dried and blended with the remainder ofthe corn starch and the magnesium stearate. The resulting granulation isthen compressed into tablets containing 50 mg, 100 mg and 300 mg of theNK-1 receptor antagonist per tablet.

We claim:
 1. A method for the treatment or prevention of a substance usedisorder, which method comprises the oral administration to a patient inneed thereof of an effective amount of an orally active, CNS-penetrantNK-1 receptor antagonist wherein the NK-1 receptor antagonist is longacting as determined by its ability to inhibit NK-1 receptoragonist-induced foot-tapping in the gerbil with an ID₅₀ less than orequal to 10 mg/kg i.v. after 24 hour pre-treatment.
 2. A methodaccording to claim 1 wherein the NK-1 receptor antagonistis:2-(S)-(3,5-bis(trifluoromethyl)benzyloxy)-3(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)-3-(S)-phenyl-morpholine;2-(S)-(3,5-bis(trifluoromethyl)benzyloxy)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)-3-(S)-phenyl-morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(5-(N,N-dimethylamino)methyl-1,2,3-triazol-4-yl)methyl-3-(S)-phenylmorpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(5-(N,N-dimethylamino)methyl-1,2,3-triazol-4-yl)methyl-3-(S)-(4-fluorophenyl)morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(4-monophosphoryl-5-oxo-1H-1,2,4-triazolo)methyl)morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(1-monophosphoryl-5-oxo-1H-1,2,4-triazolo)methyl)morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(2-monophosphoryl-5-oxo-1H-1,2,4-triazolo)methyl)morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxyphosphoryl-1H-1,2,4-triazolo)methyl)morpholine;2-(S)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(1-monophosphoryl-5-oxo-4H-1,2,4-triazolo)methyl)morpholine;2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-4-(4-N,N-dimethylaminobut-2-yn-yl)-3-(S)-(4-fluorophenyl)morpholine;(3S,5R,6S)-3-2-cyclopropoxy-5-(trifluoromethoxy)phenyl-1-oxa-7-aza-spiro 4.5!decane;(3R,5R,6S)-3-2-cyclopropoxy-5-(trifluoromethoxy)phenyl-1-oxa-7-aza-spiro 4.5!decane;(±)-(2R3R,2S3S)-N-{2-cyclopropoxy-5-(trifluoromethoxy)phenyl!methyl}-2-phenylpiperidin-3-amine;ora pharmaceutically acceptable salt thereof.
 3. A method according toclaim 1 wherein the substance use disorders are substance dependence orabuse with or without physiological dependence.
 4. A method according toclaim 1 wherein the substance associated with the disorders is selectedfrom: alcohol, amphetamines or amphetamine-like substances, caffeine,cannabis, cocaine, hallucinogens, inhalants, nicotine, opioids,phencyclidine or phencyclidine-like compounds, sedative-hypnotics orbenzodiazepines, and combinations of the above.
 5. A method according toclaim 1 wherein the substance use disorders are selected from drugwithdrawal disorders; alcohol withdrawal delirium; amphetaminewithdrawal; cocaine withdrawal; nicotine withdrawal; opioid withdrawal;sedative, hypnotic or anxiolytic withdrawal with or without peripheraldisturbances; sedative, hypnotic or anxiolytic withdrawal delirium; andwithdrawal symptoms due to addictive substances.
 6. A method accordingto claim 1 wherein the substance use disorders are selected fromsubstance-induced anxiety disorder with onset during withdrawal;substance-induced mood disorder with onset during withdrawal; andsubstance-induced sleep disorder with onset during withdrawal.